LOCUS       sequence-373985-        3139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of anti-CRISPR protein AcrVIB1 in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3139)
  AUTHORS   Wandera KG, Alkhnbashi OS, Bassett HVI, Mitrofanov A, Hauns S, Migur
            A, Backofen R, Beisel CL
  TITLE     Anti-CRISPR prediction using deep learning reveals an inhibitor of 
            Cas13b nucleases.
  JOURNAL   Mol Cell. 2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003.
  PUBMED    35649413
REFERENCE   2  (bases 1 to 3139)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003."
COMMENT     Sequence Label: pAcrVIB1-cloDF-kan
FEATURES             Location/Qualifiers
     source          1..3139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     271..290
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      432..526
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1222..1960
                     /direction=RIGHT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     terminator      complement(1980..2066)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2427..2449
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(2491..2525)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(2549..2567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(2746..3139,1..401)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2800..2819)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
       61 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
      121 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
      181 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
      241 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
      301 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
      361 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
      421 gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
      481 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa gcactaggga
      541 cagtaagacg ggtaagcctg ttgatgatac cgctgcctta ctgggtgcat tagccagtct
      601 gaatgacctg tcacgggata atccgaagtg gtcagactgg aaaatcagag ggcaggaact
      661 gctgaacagc aaaaagtcag atagcaccac atagcagacc cgccataaaa cgccctgaga
      721 agcccgtgac gggcttttct tgtattatgg gtagtttcct tgcatgaatc cataaaaggc
      781 gcctgtagtg ccatttaccc ccattcactg ccagagccgt gagcgcagcg aactgaatgt
      841 cacgaaaaag acagcgactc aggtgcctga tggtcggaga caaaaggaat attcagcgat
      901 ttgcccgagc ttgcgagggt gctacttaag cctttagggt tttaaggtct gttttgtaga
      961 ggagcaaaca gcgtttgcga catccttttg taatactgcg gaactgacta aagtagtgag
     1021 ttatacacag ggctgggatc tattcttttt atcttttttt attctttctt tattctataa
     1081 attataacca cttgaatata aacaaaaaaa acacacaaag gtctagcgga atttacagag
     1141 ggtctagcag aatttacaag ttttccagca aaggtctagc agaatttaca gatacccaca
     1201 actcaaagga aaaggactag tgatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1261 aatcttttgc cctgtaaacg aaaaaaccac ctggggaggt ggtttgatcg aaggttaagt
     1321 cagttgggga actgcttaac cgtggtaact ggctttcgca gagcacagca accaaatctg
     1381 tccttccagt gtagccggac tttggcgcac acttcaagag caaccgcgtg tttagctaaa
     1441 caaatcctct gcgaactccc agttaccaat ggctgctgcc agtggcgttt taccgtgctt
     1501 ttccgggttg gactcaagtg aacagttacc ggataaggcg cagcagtcgg gctgaacggg
     1561 gagttcttgc ttacagccca gcttggagcg aacgacctac accgagccga gataccagtg
     1621 tgtgagctat gagaaagcgc cacacttccc gtaagggaga aaggcggaac aggtatccgg
     1681 taaacggcag ggtcggaaca ggagagcgca agagggagcg acccgccgga aacggtgggg
     1741 atctttaagt cctgtcgggt ttcgcccgta ctgtcagatt catggttgag cctcacggct
     1801 cccacagatg caccggaaaa gcgtctgttt atgtgaactc tggcaggagg gcggagccta
     1861 tggaaaaacg ccaccggcgc ggccctgctg ttttgcctca catgttagtc ccctgcttat
     1921 ccacggaatc tgtgggtaac tttgtatgtg tccgcagcgc cctaggtcta gggcggcgga
     1981 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     2041 ttcgactgag cctttcgttt tatttgatgc ctttacttga gtgtagttaa gtcgcctaat
     2101 gacaacactc cacaaaggat cttgtcgctt tcacccgtga agatttcccc ttcaaaggtt
     2161 tgcaactcaa agtagctatt atcgatgtta ccccagcgtt tcgagtagat cgtgaagttt
     2221 tgcattacat cgtcctcatc aatcttcaag tagctgtaaa tgatttccac gtactcatcc
     2281 ttgtgtacca gaatgtgcat gaacccatcg tccgtatccg tacgttcgga tgagagttgg
     2341 attgcagtgg ctttcttgtt gttcaaaagc cactgggcaa tctcttcacc cttaagcttg
     2401 gacaggtcca ggtccttcat ggtatatctc cttcttaaag ttaaacaaaa ttattgctag
     2461 ccctcctgag ctcgaattcg gatcctagag gctagcatta tacctaggac tgagctagct
     2521 gtcaagaact cgaggtgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2581 gtcatgataa taatggtttc ttagacgtcg gaattgccag ctggggcgcc ctctggtaag
     2641 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     2701 aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2761 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2821 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2881 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2941 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3001 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3061 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3121 cttgatccgg ctacctgcc
//
